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1 PROCEEDINGS

2 The main objective of the international conference on Integrated biological networks driving disease outcomes is to establish transformative education and research collaborations in infectious diseases and cancer through a partnership between the University of California, Davis, USA and the Brazilian Research Network that is supported by the CNPq, CAPES and FAPEMIG. The conference will connect researchers with diverse expertise in immunology, microbiology, molecular medicine, nanotechnology and plant biotechnology to identify scientific expertise, resources and opportunities for defining the biological networks driving disease outcomes and developing new biomarkers and sensor platforms for the detection and management of infectious diseases and cancer. Luiz Ricardo Goulart Filho and Satya Dandekar Coordinators

7 NANOBIOTECHNOLOGICAL APPROACHES FOR DISEASE DIAGNOSTICS AND THERAPEUTICS. Luiz R. Goulart, PhD Universidade Federal de Uberlândia (UFU), Instituto de Genética e Bioquímica, Laboratório de Nanobiotecnologia, Campus Umuarama, Bloco 2E, Sala 248. CEP: , Uberlândia, MG, Brasil. Phone: (55+34) Fax: (55+34) ext The discovery of biomarkers for diagnostic and therapeutic purposes is the main focus of my research group, which also include the development of nanotechnologies for diagnostics, using electrochemistry and biophotonics, in association with several Brazilian and international groups. The identification of novel biomarkers; DNA/RNA aptamers, bioactive peptides and antibody fragments (scfv and Fab), are based on combinatorial technologies, including aptamer selection and Phage Display, which are associated with molecular, immunological, and imaging strategies coupled to in vitro and in vivo functional assays. Because of the transversal nature of the research group, our current findings have generated 18 patents, and several important publications in many different diseases and diagnostic platforms (electrochemistry, latex immunoagglutination, biophotonics, and magic-size quantum dots). The most exciting results have led us to the development of novel biomarkers (peptide and antibodies) for leprosy, tuberculosis, malaria, cutaneous and visceral leishmaniasis, chagas disease, bovine brucellosis, bovine anaplasmosis, and cancer (prostate, breast, mouth, and thyroid), and epitope-based vaccines for brucellosis, anaplasmosis, leishmaniasis, and bovine ticks. Currently, we have join a research effort between UCDavis and VRC-NIAID-NIH to develop a chimeric-epitope immunogen (UFU) coupled to a new virus-like particle (UCDavis), which is part of a recent UCDavis/UFU agreement. We have also shown how the cell-signaling molecule, Annexin A1, is expressed during acute and chronic HIV/SIV infection and implications of that on the disease progression. Our ultimate goal is to solve major disease hurdles, specifically on neglected tropical diseases and chronic diseases, using an arsenal of technologies, which is also the major objective of the Brazilian Nanobiotechnology Research Network, funded by CAPES and FAPEMIG agencies. Multiple technologies, such as genomics, proteomics, immunology, tissue and cell culture, immunohistochemistry, confocal microscopy, in vivo imaging, electrochemistry, atomic force microscopy, and magic-size quantum dot syntheses, are employed in a systems biology approach, in order to explain factors that favor a disease outcome.

8 GLOBAL CHALLENGES IN HIV DISEASE. CLUES TO HIV PATHOGENESIS: GUT MUCOSAL BARRIERS, GERMS AND HIV Satya Dandekar Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis. The persistence of the human immunodeficiency virus (HIV) reservoirs and incomplete resolution of chronic immune activation during HIV infection pose major hurdles in achieving complete immune recovery despite long-term anti-viral therapy. The human gastrointestinal (GI) tract harbors >80% of the immune cells in the body and also hosts 10 times more commensal bacteria than the total number of cells in the body. The immune cells are essential for protection against pathogens yet uncontrolled immune activation can cause chronic inflammatory diseases. The interplay of the gut immune system with pathogens and commensal microbiota shapes the integrity and protection to the gut epithelial barrier and immune cells that in turn controls inflammation. Unresolved inflammation contributes to tissue injury, changes in the gut microbiota and inflammatory diseases. We performed immunologic, pathologic and genomic analyses of the gut mucosal immune responses and analysis of viral genomic diversity to investigate the role of gut mucosa in HIV pathogenesis. Gut lymphoid tissue is an early target of HIV and a site for severe loss of T helper cells and disruption of epithelial barriers. Impaired gut mucosal defenses against pathogens and dysfunction in the epithelial barrier contribute to microbial translocation and chronic immune activation. A better understanding of the host-virus interactions in the gut mucosal microenvironment and their impact on anti-microbial defenses will be important to gain insights into HIV pathogenesis and to identify targets for the development of vaccine and therapeutic interventions.

9 MOLECULAR, BIOCHEMICAL AND GENOMIC DISSECTION OF PEST/DISEASE RESISTANCE IN FRUIT TREE CROPS Abhaya M. Dandekar, Prof. University of California Davis, Plant Sciences Department, Mail Stop 4; 1 Shields Ave; Davis CA Phone: , Fax: , My lab is particularly fascinated in both understanding and engineering resistance to disease and pests. The rapid and unabated spread of vector-borne disease within orchard and vineyard crops threatens the industry, the agriculture-based economy, and the livelihood of growers and orchard/vineyard labor. Early detection of vector-borne pathogens is an essential step for the accurate surveillance and management of vector-borne diseases of orchard crops. We are using genomics to profile host and pathogen responses to develop tools that would detect the infectious agents in the field in real time at early (primary) stages of infection with a high degree of sensitivity and specificity and that would enable rapid, early disease detection and diagnosis of catastrophic diseases like Huanglongbing (HLB), in citrus. These technologies have three anticipated outcomes: 1) Identification of very early, disease-specific biomarkers using a knowledge-base of translational genomic information on host and pathogen responses associated with early (asymptomatic) disease development; 2) Development and deployment of novel sensors that capture these and other related biomarkers and aid in pre-symptomatic disease detection and 3) the development of transgenic rootstocks that deliver therapeutics to combat disease and to sustain orchard and vineyard productivity. By combining the first two distinct approaches, it should be possible to identify and defend the crop by interdicting pathogen spread prior to the rapid expansion phase of the disease. The deployment of the third strategy will help preventing disease spread while sustaining productivity. We have developed engineered rootstocks that can deliver therapeutic proteins or small interfering RNA to block pathogenesis and to clear the pathogen. The rootstock based delivery of antimicrobial agents or disease prevention mechanisms are a viable strategy for both orchard and vineyard crops and especially for varietal grapevines that cannot be genetically modified. We have demonstrated rootstock-based delivery of antimicrobial proteins and RNA interference to combat Pierce s Disease and bacterial crown gall disease successfully. Engineered rootstocks represent a viable genetic strategy to sustainably manage the spread of disease and pests with global climate change and to reduce the use of chemical pesticides.

10 HOW THE HOST RESPONSE FEEDS SALMONELLA Andreas J. Bäumler Department of Medical Microbiology and Immunology, School of Medicine, University of California at Davis, Davis, CA The intestine is host to a diverse bacterial community whose structure, at the phylum level, is maintained through unknown mechanisms. Acute inflammation triggered by enteric pathogens, such as Salmonella enterica serotype Typhimurium (S. Typhimurium), is accompanied by changes in the bacterial community structure marked by an outgrowth of the pathogen. Recent studies show that S. Typhimurium can harness benefit from the host response to edge out the beneficial bacterial species that dominate in the healthy gut. The elucidation of how S. Typhimurium alters the bacterial community structure during gastroenteritis is beginning to provide insights into mechanisms that dictate the balance between the host and its microbiota.

11 STRUCTURAL MODULARITY IN THE MOLECULAR DESIGN OF TARGETED VACCINES R Holland Cheng, PIOMS Institutional Program, Human immunodeficiency virus (HIV) Env complex is comprised of a trimer of gp120 associated with membrane-embedded gp41 trimers. Receptor association exposes a cryptic epitope for coreceptor engagement, leading to host and viral membrane fusion. Our previous work (Moscoso et al., 2011 PNAS 108:6091-6) with clade C Env trimers provided us with a base for comparison with native Env conformation, in order to elucidate any quaternary differences in trimer morphology, epitope location, and occlusion of antigenic regions. Such a targeted structural survey of HIV Env in complex with ligands such as primary receptor CD4, the broadly neutralizing antibody VRC01, as well as trimer conformation-recognizing antibodies such as PG16 reveals pertinent information regarding the location of these epitopes. The spatial configuration of selected ligand footprints relative to the CD4 and coreceptor binding sites provides insight of mechanism in various metastable conformations transpired, where novel, potentially quaternary neutralizing epitope (QNE) may be exposed to elicit broadly neutralizing antibodies. Additionally, the use gp140-s-s-cd4m and single chain gp140-cd4m constructs, in the discovery of immunogens with novel QNE, eschews any concerns of auto-immune reactions by only implementing the short, 27-residue CD4m sequence into the Env sequence, in order to minimally impact proper protein folding.

12 CANCER MOLECULAR BIOLOGY A MEANS TO IMPROVE CANCER PATIENT OUTCOMES Ralph de Vere White, M.D. University of California, Davis (UCD), UCD Comprehensive Cancer Center, 4501 X Street #3003, Sacramento, Ca., Phone: (916) Fax (916) The UC Davis Comprehensive Cancer Center (UCDCCC) consists of 308 members, 45% of who are outside of the medical school, organized into 6 programs, 5 initiatives and 10 shared resources. In the past decade the centers funding has grown from $42 million to $112 million (USD) yearly. Presently, a major focus is to integrate molecular/genetic information into patient care, pharmacogenetics, drug discovery with biomarker development as a strategy to overcome the constrictions in translational research to ultimately improve health. To accomplish this, we have focused on delivering patient cancer care as informed by their tumors molecular profile. Regrettably, present molecular knowledge is insufficient to control cancer. To remedy this, we are using two platforms with the aim of finding therapeutic targets in individual cancers that will allow repurposing of therapies used presently in other cancers or other diseases. We believe, while this will not replace the need for new drug discovery, it will allow for improvement in patient outcomes in a much shorter time frame. We will focus on vertical genome to metabolomic analysis of selected tumors for this drug repurposing program, in two platforms. The first is integrating studies with our Veterinary school, which has 8,000 visits, mostly dogs with spontaneous tumors every year. Second, is the acquisition of tumors from biopsies and in the operating rooms, which are then transplanted, un-manipulated, into NSG mice. Since these mice lack natural killer cells, tumor take is excellent. Presently, the major molecular drivers found in the patient tumors are faithfully reproduced in the NSG mouse model, for at least 6 passages. Examples of how these models are being used in lung, prostate, and urothelial cancer to improve patient outcomes will be presented.

13 HOST-PATHOGEN INTERACTIONS OF BRUCELLA LEADING TO CHRONIC INFECTION Renée M. Tsolis, Ph.D. Medical Microbiology and Immunology, University of California, Davis Phone: Fax: Brucella spp are zoonotic pathogens that cause one of the most widespread yet neglected zoonotic diseases, namely brucellosis. These bacteria cause chonic infections both in the zoonotic reservoir host species (primarily cattle and goats), and in humans, impacting both agriculture and human health. Persistent infection is associated with the ability of Brucella to persist within cells of the mononuclear phagocyte system. Our laboratory, in collaboration with Dr. Renato Santos group at UFMG, is dissecting the host and pathogen factors that allow this infection to progress to a chronic disease and impede an effective and sterilizing immune response. One of the bacterial factors, the Type IV secretion system (T4SS), contributes both to survival within phagocytes and persistent infection. Until recently, the substrates of this secretion system were unknown. We have identified one of the translocated substrates, named virb-coregulated effector C (VceC), which is co-expressed with the T4SS structural genes. Our results showed that VceC played only a minor role in persistence in the mouse infection model, but induced an inflammatory response in infected macrophages. Ectopically expressed VceC targeted to the endoplasmic reticulum (ER), the intracellular site of B. abortus replication, where it interacted with the ER chaperone BiP/GRP78 and led to disruption of ER structure. B. abortus infection led to a modest induction of ER stress, which was partially dependent on VceC. Our results suggest that VceC may contribute to the T4SS-dependent inflammatory response via induction of ER stress. This inflammatory response could contribute to bacterial persistence via stimulation of granulomatous inflammation in the spleen, or possibly to transmission in the zoonotic reservoir via abortion. To address the role of these host factors in the biology of zoonotic transmission and persistent disease, studies in the zoonotic host reservoir are urgently needed, which will necessitate collaboration between the fields of veterinary science and reproduction, immunology and microbiology.

14 NANOSTRUCTURED CERAMICS DEVICES AS POTENTIAL BIOLOGICAL SENSORS Ricardo H. R. Castro, PhD Division of Materials Science,Peter A. Rock Thermochemistry of Nanoceramics Laboratory, University of California, Davis. The macro-properties of nanostructured materials are strongly dependent on their surface features. This is because a large fraction of their atoms is within the interface region, suggesting that changes to the surface chemistry can significantly influence, for instance, their electrical properties. This influence has been exploited to develop highly sensitive and selective sensors for gas phase molecules using engineered nanostructures. In this talk, we show some recent advances on the gas sensing capabilities of devices built using tin dioxide and carbon nanotubes. We show that the reversible absorption of specific gases by the sensor can change the conductivity of the nanoparticulate film, generating an unambiguous signal for toxicity control. We suggest the possibility of using similar setups for biological applications by highlighting successful results from the literature.

15 HOST-PARASITE RELATIONSHIP OF APICOMPLEXAN PROTOZOA Tiago W. P. Mineo, PhD Universidade Federal de Uberlândia, Instituto de Ciências Biomédicas, Laboratório de Imunoparasitologia Dr. Mário Endsfeldz Camargo, Campus Umuarama, Bloco 4C, Sala 4C01. CEP , Uberlândia, MG, Brasil. Phone: (55 34) Fax: (55 34) Neospora caninum and Toxoplasma gondii are closely-related apicomplexan parasites with worldwide distribution, and both are able to infect a wide range of warm-blooded hosts. The mechanisms underlying host resistance against these pathogens remain unclear and have been the subject of study by our group. In order to successfully obtain efficient biotechnological tools for the diagnostic, therapy and prophylaxis against both infections, we have worked on the advancement of knowledge in the fields of epidemiology, differential host and parasite protein expression, peptide immunomapping, innate and adaptive signaling pathways, adjuvant selection, as well as in vivo and in vitro models.

16 DEVELOPMENT OF BIOSENSORS APPLIED IN THE DIAGNOSIS OF HUMAN DISEASES Ana Graci Brito Madurro 1 and João Marcos Madurro 2 Laboratory of Polymer Films and Nanotechnology LAFIP-NANOTEC. Institute of Chemistry, Federal University of Uberlândia Brazil. Laboratory of Biomaterials LABIOMAT. Institute of Genetics and Biochemistry, Federal University. of Uberlândia Brazil. The current scenario in the detection of infectious and inflammatory diseases requires new platforms able to identify them in the early stages of infection. Polymer matrices have good thermal and electrochemical stability with storage time and serve as excellent sensitizers for biosensors development. DNA probes, antigens, antibodies and enzymes are able to recognize specific targets and are used by our research group, focusing on the detection of infectious and inflammatory diseases such as acute myocardial infarction, hepatitis C, breast cancer, ovarian cancer, prostate cancer, leprosy, dengue, leishmaniasis, meningitis and others, in partnership with Brazilian research groups. The electrochemistry is the principal technique used for preparation of the polymers and detection of the targets with the biosensors produced, however other methods are used to validate the results. The selectivity, sensitivity and stability are tested in order to optimize the electrochemical biosensors. The results obtained using synthetic and real samples have indicated that these devices are versatile and interesting for use in hospitals, clinics, blood banks and testing in-house. Our research group is grateful for the financial support from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) and Coordenação de Aperfeiçoamento de Pessoal Nível Superior (CAPES).

17 BIOMARKERS FOR LEPROSY SURVEILLANCE: FINDING NEW CASES BY NOVEL APPROACHES Claudio Guedes Salgado, MD, PhD Universidade Federal do Pará (UFPA), Instituto de Ciências Biológicas, Laboratório de Dermato- Imunologia, Marituba, Pará, Brasil. CEP: Tel: The occurrence of leprosy among children is correlated with recent disease and active foci of transmission in the community. We performed clinical and serological examinations of 1592 randomly selected school children (SC) during a cross-sectional survey in 8 hyperendemic municipalities of the Brazilian Amazon region. Sixty-three (4%) SC, with a mean age of 13.3 years (SD = 2.6), were diagnosed with leprosy, and 777 (48.8%) were seropositive to anti-pgl-i. Additionally, we evaluated 256 household contacts of those students diagnosed with leprosy: 24 (9.4%) of them were also diagnosed with leprosy and 107 (41.8%) were seropositive. Seroprevalence was significantly higher among girls, students from urban areas and students from public schools (p <0.0001). Forty-five (71.4%) new cases detected among SC were classified as paucibacillary, and 59 (93.6%) did not demonstrate any degree of physical disability at diagnosis. These results suggest a high rate of undiagnosed leprosy and subclinical infection among children in the Amazon region. The advantages of school surveys in hyperendemic areas are finding early cases of leprosy with no physical disabilities, preventing the spread of the infection in the community and breaking the chain of transmission. A new method for finding new cases was developed by our group using anti-pgl-i results on visited municipalities. At Oriximiná, Northwest Pará, we found 23 new cases among anti-pgl-i + SC and their contacts, while only 7 were found on the anti-pgl-i negative group. Using the same approach, examining only anti-pgl-i+ SC and their contacts, or anti-pgl-i+ contacts of leprosy cases, our group found more than 70 new cases in one week work at Castanhal, a medium city 65 Km from Belém, capital of Pará State. We are now on a joint effort with Colorado State University, to find if new biomarkers, as LID-I, LAM and others may be useful to increase our capability on finding new cases of leprosy.

18 GENETIC ENGINEERING APPROACHES IN THE DEVELOPMENT OF IMMUNOMODULATOR BIOACTIVE PEPTIDE AND IN CONSTRUCTION OF TRANSGENIC MOSQUITOES Carlos Ueira-Vieira, PhD Universidade Federal de Uberlândia (UFU), Instituto de Genética e Bioquímica, Laboratório de Nanobiotecnologia, Campus Umuarama, Bloco 2E, Sala 248. CEP: , Uberlândia, MG, Brasil. Phone: (55+34) ext.27. Transforming Growth Factor-1 (TGF-β1), cytokine 10 (IL-10) and tumor necrosis factor (TNF-α) are polypeptides that plays crucial roles in embryogenesis, tissue development, carcinogenesis and immune regulation. They are involved in suppression of inflammatory response in the pathogenesis of autoimmune diseases (Juvenile Idiopathic Arthritis, multiple sclerosis, diabetes), through the activation of regulatory T cells (Tregs). In recent years, mapping affinity-selected peptides screened from a random phage display library to the native epitope has become popular in epitope prediction. These peptides, also known as mimotopes, share the similar structure and function with the corresponding native epitopes. Our research groups have isolated same bioactive mimetics peptides obtained by Phage Display to TGF- β1, IL-10 and TNF-α. Stimulation of human PBMC in vitro showed that same mimetics cytokines bioactive peptides have the agonist function and other as antagonists which can be considered potential targets for development immunomodulators drug. Our group developed also peptide that can binding in specific inflamed tissues, we used murine model of rheumatoid arthritis (DBA1/)1 and we found also specific peptides that binding in blood vessel. Theses peptides can be used for imaging diagnostic and drug delivery to specific inflammatory site to minimize systemic effects and immunosuppressive drugs. Our group also uses genetic engineering to produce transgenic animals. We are using a new silencing proteins technology to construct genetically modified mosquitoes where dengue virus is unable to replicate. Financial Support: CNPq, FAPEMIG, CAPES and UFU

19 A JOURNEY INTO TRYPANOSOMA CRUZI-HOST INTERACTION Claudio Vieira da Silva, Adele Aud Rodrigues, Fabrício Castro Machado, Thaise Lara Teixeira, Marlus Alves dos Santos Universidade Federal de Uberlândia, Instituto de Ciências Biomédicas, Laboratório de Tripanosomatídeos, Bloco 6T sala 07, Uberlândia, MG, Brazil. Chagas disease is caused by the kinetoplastid protozoan parasite Trypanosoma cruzi. One of our greatest challenge in the laboratory was to understand why amastigotes from T. cruzi G strain are highly infective in vitro and do not contribute for a patent in vivo infection. Our in vitro studies demonstrated the first evidence that IFN-γ would be associated to the in vivo low virulence of G strain. After intraperitoneal amastigotes inoculation in wild-type and knockout mice for TNF-α, Nod2, Myd88, inos, IL-12p40, IL-18, CD4, CD8 and IFN-γ we found that the latter is crucial for controlling infection by G strain amastigotes. Considering parasite protein associated to cell invasion it is worth mentioning that T. cruzi P21 was recently characterized and its activity during parasite internalization in host cell has been studied. We have observed that P21-His 6 upregulates phagocytosis via CXCR4 chemokine receptor signaling pathways. Also, it showed chemotactic activity both, in vitro and in vivo. Host cell membrane proteins pose as important ligands to T. cruzi infective forms. In this sense, we have studied the role of Galectin-3 during T. cruzi cell invasion and intracellular traffic. We have demonstrated that extracellular amastigotes from T. cruzi were associated with a galectin-3 containing structure at times related to the vacuole lysis in mouse embryonic fibroblasts. Moreover, we have found that somehow Galectin-3 controls amastigote intracellular multiplication. The chemotherapy used in treatment of Chagas disease is considered ineffective and has serious side effects. We observed that extracts derived from plants belonging to the genus Ruta, Ageratum and Tabebuia showed high toxicity against T. cruzi tripomastigotes and intracellular amastigotes, impairing parasite multiplication. Financial Support: FAPEMIG, CAPES and CNPq.

20 SMALL RNA-REGULATED PATHWAYS ASSOCIATED WITH VEGETATIVE AND REPRODUCTIVE ORGAN DEVELOPMENT IN PLANTS Fabio TS Nogueira, PhD Instituto de Biociencias, UNESP, Botucatu, SP, Brazil. Phone: (55+14) Plants, unlike animals, are extremely plastic in responding to changes in the environment and they can shape their own development. Plants are capable to generate new tissues/organs throughout their entire life cycle. The establishment and development of such tissues/organs depend upon several physiological and molecular processes that take place inside the cells. Our research group is interested in studying the formation of vegetative and reproductive organs as well as how plants shape these organs in response to the environment. More specifically, we are interested in understanding the molecular mechanisms that underline the formation of these organs. Several genetic players are involved in such mechanisms, including transcription factors, epigenetic factors and non-coding RNAs. Small non-coding RNAs (srnas) of nt in size regulate transcriptionally and posttranscriptionally gene expression, shaping the transcriptome and the proteome of the cells. We have identified srnas associated with sugarcane (an important Brazilian biofuel crop) axillary bud dormancy and development. We identified for example an emerging new class of srnas called trna-derived fragments or trfs that may have roles in cell differentiation. We are also studying the roles of specific micrornaregulated pathways in tomato fruit and leaf development. By using a diverse set of molecular and genetic approaches, we intend to understand the roles of this complex class of molecules in plant development that may contribute to the generation of plant biomass.

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